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Evaluation of agricultural effluents and irrigation water as sources of antimicrobial resistant Escherichia coli

Romanis, Marco (2013-12)

Thesis (MSc Food Sc)--Stellenbosch University, 2013.

Thesis

ENGLISH ABSTRACT: Food-borne disease outbreaks caused by Escherichia coli have been linked to the use of faecally-polluted irrigation waters. Thus the overall aim of this research was to evaluate irrigation water and agricultural effluents as sources of antibiotic resistant E. coli in the Western Cape. The aim of the first study was to enumerate and characterise E. coli present in irrigation water and in potential contamination sources. Maximum total coliform and E. coli counts for irrigation sites was log 7.862 and log 5.364 MPN.100 mL-1, respectively. Five out of seven irrigation sites had E. coli counts exceeding national and international guidelines for ‘safe’ irrigation water (<1 000 counts.100 mL-1), making it unsafe for the irrigation of fresh produce.
In this study, 46.6% of the E. coli strains were characterised in phylogenetic group B1. It has been shown that E. coli in group B1 have the ability to survive and persist in the external environment. Group B1 was also the most common group among isolates from irrigation sites (79.4%), while isolates from environmental sites grouped mainly in group A0 (54.1%). It was concluded that the wide variation of E. coli types present in irrigation water is a concern that should be further investigated. This raises human health implications since the increased exposure to faecal organisms increases the risk of food-borne outbreaks.
The E. coli isolates (n = 120) and the marker (n = 37) and reference strains (n = 6), were evaluated for antibiotic resistance to seven medically-important antibiotics from different classes using the Kirby-Bauer disc diffusion method. Thirty-five strains (35/163 = 21.5%) exhibited resistance to one or more antibiotics. Piggery effluent was found to harbour the most antibiotic resistant E. coli isolates (9/35 = 25.7%). Among the resistant E. coli strains, the highest occurrence of antibiotic resistance was to trimethoprim (2.5 μg) (68.6%), tetracycline (30 μg) (57.1%), ampicillin (10 μg) (45.7%) and chloramphenicol (30 μg) (34.3%). Seventy-four percent (26/35) exhibited multiple antibiotic resistances to two or more antibiotics.
The antibiotic resistant E. coli strains were evaluated for the presence of pathotypes using Polymerase Chain Reaction analysis to detect Intestinal Pathogenic E. coli (InPEC) and Extra-intestinal Pathogenic E. coli (ExPEC). Five InPEC strains were characterised as four Entero-Pathogenic E. coli (EPEC) strains resistant to three or four antibiotics and one Entero-Aggregative E. coli (EAEC) strain resistant to trimethoprim. The antibiotic resistant EAEC strain also possessed the ExPEC-related gene iutA. Two E. coli isolated from the Mosselbank River were both resistant to chloramphenicol and trimethoprim and also possessed the ExPEC-related gene iutA. It was concluded that the diverse antibiotic resistances of E. coli pathotypes present in irrigation water is a concern that should be further investigated.